17alpha, 21-oxido derivatives of cortisone and hydrocortisone



United States Patent 2,836,593 17,21-0XIDO DERIVATIVES OF CORTISONE AND HYDROCORTISONE Roger E. Beyler, Westfield, and Frances Hoffman,

Newark, N. 1., assignors to Merck & (30., Inc., Rahw'ay, N. J., a corporation of New'Jersey No Drawing. Application April 29, E57 Serial No. 655,526 5 Claims. (Cl. 260239.55)

This invention relates to adrenal cortical hormones having cortisone-like anti-inflammatory activity and more particularly to the 17a,21-oxido derivatives of cortisone and hydrocortisone.

The compounds of the present invention have the general formula CHa I C-GH:

2,836,593 Patented May 27, 1958 responding 2l-fiuoro compound (21-fiuoro-4-pregnene- 17u-ol-3,11,20-trione IV, or 21-fluoro-4-pregnene-11b,17adiol-3,20-dione) is not the only or even the predominant steroid product of this reaction. Instead, a substantial quantity of II is converted to l7ot,2l-oxido-4-pregnene- 3,11,20-trione (III). Similarly, hydrocortisone methanesulfonate is convertted in large part to 17u,21-oxido-4- pregnene-l1,B-ol-3,20-dione. Compounds III and IV (or the corresponding l7a,21-oxido and 21-fiuoro derivatives of hydrocortisone) can be separated chromatographically. For example, III and IV can be extracted from the reaction product mixture with a suitable organic solvent such as methylene chloride, chloroform, ethyl acetate, or benzene, chromatographed, and the adsorbent developed with a suitable solvent combination. One such combination consists of ether and chloroform, from which the product (IV or hydrocortisone-l7a,21-oxide) is recovered in the ether eluate.

The compounds of the present invention have cortisone- Iike anti-inflammatory activity. These compounds are also useful as intermediates for conversion to the l7a,21- oxido derivatives of'prednisone and prednisolone, which possess greater anti-inflammatory activity. This conversion may be efifected by a suitable dehydrogeuating agent such as selenium dioxide or micro-organisms such as CH CHa CHnOH CHgOSOzCHa CH CH3 CHaSOzCl I II 0 CH (H) CH2]? GH2 (E =0 l---0H O- 0- CH3 CH III dried to give 6.6 g. of product.

Bacillus sphaericus. The conversion of III to the 17u,21-

oxido derivative of prednisone (V) may be represented by the following equation:

L-CH3 170:,21-oxido 4-pregnene 1113 ol3,20-dione may be converted similarly to 17a,21-oxido-1,4-pregnadiene-113-01- 3,20-dione. W V s This invention will now be described with reference to specific examples thereof. i

7 EXAMPLE I 7 7 Preparation of 170:,21-0xide-4-pregnene-3,1LZO triOne One gramof cortisone-2l-methanesulfonate (II) and 1.75 g..of potassium fluoride weresuspended in 35 ml. 'of ethylene glycol in a 100-ml. round-bottomed flask., The

7 reaction mixture was heated in a nitrogen atmosphere for 4 /2 hours at 100 C." The mixture was then 'c'ooledin an ice bath, diluted with 100 ml. of .Waterand'extracted with two portions of 100 ml. each of methylene chloride. The extract was dried over anhydrous magnesium sulfate and the methylene chloride was then evaporated in vacuo. The residue which weighed 883 mgrwas dissolved.

in-S ml. of benzene and chromatographed'on a column containing 25 g. of acid-washed, acetone-treated alumina.

The column was eluted successively with 250 ml. of ether, 150 ml. of a 60:40'(by volume) chloroform-ether'mixture, 200 ml. of a 80:20chloroform-ether mixture and 250 ml. of chloroform. The product 17a,21-oxido-4-preg-' 3 i nene-3,11,20-trione (HI) was recovered from the ether eluate... Yield 464 mg.; MIP. 208-2l1 C. (215 C.);

' infrared spectrum, 5.56, 5.90, 6.02, 6.19; ultraviolet spectrum: A max; 238, E% 475. 7

Analysis.'Calculated: C, 73.66%; H, 7.66%. Found:

An equivalent quantity of hydrocortisone may be converted to 170:,21 oxido 4 pregnene 11p o1 3,20 dione by the proeedure'of Example 1.

EXAMPLE 2 Conversion of 170:,21 oxz'do 4 pregnene 3,11,20 m

one (III) to 170:,21 oxido 1,4 L pregnadiene 3,11,20- trione (V) r s l.

Three hundred mg. of 1711,21 oxido 4 pregnanc- 3,11,20-t1'ione were dissolved in 21 ml. of'dry terti butanol, and 0.25 ml. of glacial acetic acid was addedif To this mixture 100 mg. of selenium dioxide were added and the. mixture refluxed under nitrogen for .24 hours. I: The

supernatant liquid was decanted from the selenium and concentrated to dryness under vacuum. The residue was suspended in ml. of water, extracted with 10 ml. of ethyl acetate, the extract separated, and the water layer extracted twice again with 10 ml. of ethyl acetate. ethyl acetate extracts were combined, washed with dilute The , hydrochloric acid and aqueoussodium bicarbonate,.and The ethyl acetate was which 'c omprises' combining the, coiresponding ZO-keto-" I appended claims.

dried over magnesium sulfate. distilled ofi. The residue was dissolved in 10 ml. of benzene, chromatographed on 10 g. of acetoneactivated, acid-washed alumina, and eluted with mixtures of petroleum ether and ether, starting with a 1:1 mixture of petroleum ether and ether, gradually increasing the amount of ether and finally eluting with pure ether. The 1711,21- oxido 1,4 pregnadiene 3,11,20 trione product was recovered from the ether eluate and recrystallized from acetone-petroleum ether; Ml- "PL242 to 244 C;

The procedure of ExamplejZjcan also be usedto convert 170:,21 oxidio- 4 pregnene. 11,8 -.ol 3,20 dione to 17a,2l-oxido-1,4-pregnadiened1fl-olr3,20-dione.1 7

-. EXAMPLE 3, ,7 2 V A nutrient medium having thefollowing composition:

KOH to adjustpH to 6.5. e 1 r I1 was'sterilized by autoclaving at 120 C."and intrdduced in 25-ml. portions into'1 25-m l. shake flasks. A culture I of Bacillus sphaericus (MB-880) was grown on the above nutrient medium for 24 hours" at 28 C. .in the shake flasks, which were agitated on a rotary shaker at 220 R. P. M. Then Sirng; of 17a,21-oxido-4-pregnene-l lfl-ol- 1 3,20-dione in dimethylformamide solution, having a concentration of 100 mg./ml., was added to each flask, and

incubation was continued for an additional 24 hours under the same conditions. The whole broth was then extracted with an equal volume of ethyl acetate in three equal portions, and the extracts were combined and concentrated to 3 ml. The concentrate was spotted on paper and developed in the system benzene-formamide according to the procedure of-Zaifaroni'et al., jScience"11 1,i6 (1950). The running rate of the material was identical with that of an authentic sample of 17a,21-oxido-1,4preg-' methanol wasevaporated, and the residue dissolved in' sulfuric acid. Analysis made-at the end Of'jiWQ hours showed maxima at 235; 265, 287, and 383 my.

While this invention has been described with-reference to specific embodiments thereof, it is understood that 'these embodiments are illustrative and that the scope of this invention is measured only by the scope of the What is claimed 'isi r t 1. A process for producing a compound having the general formula r r r item,

- 7 V ii-om of the group where R is selected from the group consisting of 0:;

and

17a,21-dihydroxy compound with methanesulfonyl chlohydroxy-Zl-methanesulfonate, combining the 20-keto-17ahydroxy-Zl-methanesulfonate at elevated temperature with an alkali metal fluoride, and separating the resulting l7a,2l-oxido compound from the reaction mixture.

2. A process for producing a compound of the group having the general formula O -CH:

where R is selected from the group consisting of 0.: and

which comprises combining the corresponding ZO-keto- 17a,21-dihydroxy compound with methanesulfonyl chloride, thereby forming the corresponding 20-keto-17rxhydroxy 21 methanesulfonate, combining the 20- keto-17a-hydroxy-2l-methanesulfonate at elevated temperature with an alkali metal fluoride, dissolving the steroids in the reaction mixture in an organic solvent, chromatographing the resulting solution, and recovering a fraction containing a major amount of 17a,21-0xido unsaturated pregnane compound.

3. A process for producing the compound having the formula 0 i i-CH2 which comprises combining cortisone with methane-sulfcnyl chloride, thereby forming A -pregnene-17aol3,11, ZO-trione-Zl-methanesulfonate, combining said compound at elevated temperature with an alkali metal fluoride, and separating the resulting 17a,2l-oxido-Aflpregnene-Ll1, ZO-trione.

4. A process for producing the compound having the fionnula O i l-CH2 No references cited. 

1. A PROCESS FOR PRODUCING A COMPOUND OF THE GROUP HAVING THE GENERAL FORMULA 